- Email: [email protected]
Antioxidative effect of methanolic extract of Sargassum thunbergii in macrophage RAW264.7 and HL60
S538
Abstracts / Journal of Biotechnology 136S (2008) S527–S540
VI1-P-035 A novel allergen in the soft coral which resembles to green fluorescent protein families Yuko Kato 1,∗ , Mitsuru Jimbo, Tatsuya Takahashi 1 , Sachiko Matsuhashi, Seiko Shigeta, Yukimitsu Imahara, Reiko Onizuka, Hisao Kamiya 1 Department of Environment Engineering, Tohwa University, Minamiku, Fukuoka-shi 815-8510, Japan
derivatives was investigated against hydroxyl and alkyl radicals using electron spin resonance method (ESR), and intracellular hydroxygen peroxide in live cells using 2 ,7 -dichlorofluorescein (DCF) intensity (Mendis et al., 2007; Engelmann et al., 2005). Three COS derivatives were synthezied and had antioxidant activity in ascending order cinnamyl COS, coumaryl COS and ferulyl COS. Furthermore, they were more soluble in water and non-cytotoxic compared with phenolic acids. These results suggest that COS derivatives have antioxidant effect stronger than COS and can be applied to a lot of field in the future.
E-mail address: [email protected] (Y. Kato). Certain marine organisms have been known to cause allergic reaction such as conjunctivitis, dermatitis, and bronchial asthma among occupational fishermen or inhabitant of coastal area. Bronchial asthma among the workers engaged in spiny lobster fishing on Miyazaki Prefecture, Japan, has been reported to be such as Dendronephthya nipponica and Alcyonium gracillimum, sensu Takeda et al. 2000, a junior synonym of Scleonephthya gracillima (Takeda et al., 2000). In this study for the new allergic marine animals, S. gracillima was also found to be the causative organism for allergic reactions among the fishermen in spiny lobster fishing. The novel allergen was separated from new materials of S. gracillima extract in combination with gel filtration and column chromatography on ion exchangers by monitoring ELISA reaction and Western blotting. The major allergen was identified as a protein having a molecular mass of 28 kDa in SDS-PAGE under reduced conditions, western blotting, and ECL immune staining method (Fig. 1). The in-gel digestion of 28 kDa component using trypsin was conducted, and the peptides mixture obtained were analyzed for the sequences with a LC–ESI-MS/MS LCQ deca XP in positive ion mode. Similarity search with MASCOT in NCBInr database for the sequence of YPADLPDYFK in 28 kDa component revealed the identity with the amino acid sequence of cyan fluorescent protein in Acropora aculeus and Dendronephthya sp. (Pakhomov et al., 2004). It is of quite interest that the allergen fraction showed a characteristic fluorescent spectrum, the excitation at 298 nm and the emissions at 503 nm, 570 nm and 663 nm simultaneously.
References Pakhomov, A.A., Martynova, N.Y., Gurskaya, N.G., et al., 2004. Photoconversion of the chromophore of a fluorescent protein from Dendronephthya sp. Biochemistry (Mosc) 69, 901–908. Takeda, Y., Kawakami, H., Shigeta, S., Kamiya, H., Onizuka, R., Tanabe, K., Kato, Y., 2000. Purification of a novel allergen (SAP-1), which induces allergic asthma, from the red soft coral (Alcyonium gracillimum). Allergol. Int. 49, 213–218.
doi:10.1016/j.jbiotec.2008.07.1264 VI1-YP-026
References Engelmann, J., Volk, J., Leyhausen, G., Geurtsen, W., 2005. ROS formation and glutathione levels in human oral fibroblasts exposed to TEGDMA and camphorquinone. J. Biomed. Mater. Res. Part B: Appl. Biomater. 75B, 272–276. Jeon, Y.J., Kim, S.K., 2001. Effect of antimicrobial activity by chitosan oligosaccharide N-conjugated with asparagine. J. Microbiol. Biotechol. 11, 281–286. Mendis, E., Kim, M.M., Rajapakse, N., Kim, S.K., 2007. An in vitro cellular analysis of the radical scavenging efficacy of chitooligosaccharides. Life Sci. 80, 2118–2127.
doi:10.1016/j.jbiotec.2008.07.1265 VI1-YP-027 Antioxidative effect of methanolic extract of Sargassum thunbergii in macrophage RAW264.7 and HL60 Jung-Ae Kim 1 , Young-Sook Cho 1 , Won-Kyo Jung, Moon-Moo Kim, Se-Kwon Kim 1,∗ 1 Department of Chemistry, Pukyong National University, Busan 608737, Republic of Korea
Reactive oxygen species (ROS) are generated in all aerobic organisms. Imbalances in the production and removal of ROS have been hypothesized to play a causative role in numerous diseases such as cancer, neuronal degenerative diseases such as photoaging, atherosclerosis, and arthritis. Algae extracts containing a large amount of polyphenolic compounds have been studied as potential natural antioxidants in recent years. The aim of this study was to investigate potential antioxidant effects of marine algae on cultured macrophage RAW264.7 and HL60 cells. The brown algae Sargassun thunbergii is distributed widely coastal area of East Asia. In this study, the free radical scavenging effects of methanolic extract S. thunbergii (ST) on ROS production was investigated. The levels of intracellular ROS was measured using 2,7-dichlorofluorescindiacetate. Furthermore, membrane lipid peroxidation and protein oxidation inhibition was significantly decreased in a dose-dependent manner by treatment of ST extracts. Besides, the ST extract inhibited myeloperoxidase (MPO) activity which coverts H2 O2 into HOCl in human myeloid cell (HL60) and DNA oxidation caused by hydroxyl radical generated by Fenton reaction.
Antioxidant effect of novel chitooligosaccharides derivatives Dai-Nghiep Ngo 1,2,∗ , Moon-Moo Kim, Won-Kyo Jung, Sang-Hoon Lee, Tae-Kil Eom, Se-Kwon Kim 1 1
Marine Bioprocess Research Center, Pukyong National University, Busan 608-737, South Korea 2 Department of Biochemistry, Faculty of Biology, University of Sciences, VNU-HCMC, Ho Chi Minh City, Viet Nam E-mail address: [email protected] (D.-N. Ngo).
The object of this research is to improve antioxidant effect of chitooligosaccharides (COS) by grafting some phenolic acid such as cinnamic acid, coumaric acid and ferulic acid onto amino groups of COS (Jeon and Kim, 2001). The antioxidant effect of novel COS
References Li, C.W., Ja, A.A., In, W.I., 2005. Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts. J. Agric. Food Chem. 53, 4207–4212. Rajapakse, N., Kim, M.M., Kim, S.K., 2007. Inhibition of free radical-mediated oxidation of cellular biomolecules by carboxylated chitooligosaccharides. Bioorg. Med. Chem. 15, 997–1003.
doi:10.1016/j.jbiotec.2008.07.1266
Abstracts / Journal of Biotechnology 136S (2008) S527–S540
VI1-P-035 A novel allergen in the soft coral which resembles to green fluorescent protein families Yuko Kato 1,∗ , Mitsuru Jimbo, Tatsuya Takahashi 1 , Sachiko Matsuhashi, Seiko Shigeta, Yukimitsu Imahara, Reiko Onizuka, Hisao Kamiya 1 Department of Environment Engineering, Tohwa University, Minamiku, Fukuoka-shi 815-8510, Japan
derivatives was investigated against hydroxyl and alkyl radicals using electron spin resonance method (ESR), and intracellular hydroxygen peroxide in live cells using 2 ,7 -dichlorofluorescein (DCF) intensity (Mendis et al., 2007; Engelmann et al., 2005). Three COS derivatives were synthezied and had antioxidant activity in ascending order cinnamyl COS, coumaryl COS and ferulyl COS. Furthermore, they were more soluble in water and non-cytotoxic compared with phenolic acids. These results suggest that COS derivatives have antioxidant effect stronger than COS and can be applied to a lot of field in the future.
E-mail address: [email protected] (Y. Kato). Certain marine organisms have been known to cause allergic reaction such as conjunctivitis, dermatitis, and bronchial asthma among occupational fishermen or inhabitant of coastal area. Bronchial asthma among the workers engaged in spiny lobster fishing on Miyazaki Prefecture, Japan, has been reported to be such as Dendronephthya nipponica and Alcyonium gracillimum, sensu Takeda et al. 2000, a junior synonym of Scleonephthya gracillima (Takeda et al., 2000). In this study for the new allergic marine animals, S. gracillima was also found to be the causative organism for allergic reactions among the fishermen in spiny lobster fishing. The novel allergen was separated from new materials of S. gracillima extract in combination with gel filtration and column chromatography on ion exchangers by monitoring ELISA reaction and Western blotting. The major allergen was identified as a protein having a molecular mass of 28 kDa in SDS-PAGE under reduced conditions, western blotting, and ECL immune staining method (Fig. 1). The in-gel digestion of 28 kDa component using trypsin was conducted, and the peptides mixture obtained were analyzed for the sequences with a LC–ESI-MS/MS LCQ deca XP in positive ion mode. Similarity search with MASCOT in NCBInr database for the sequence of YPADLPDYFK in 28 kDa component revealed the identity with the amino acid sequence of cyan fluorescent protein in Acropora aculeus and Dendronephthya sp. (Pakhomov et al., 2004). It is of quite interest that the allergen fraction showed a characteristic fluorescent spectrum, the excitation at 298 nm and the emissions at 503 nm, 570 nm and 663 nm simultaneously.
References Pakhomov, A.A., Martynova, N.Y., Gurskaya, N.G., et al., 2004. Photoconversion of the chromophore of a fluorescent protein from Dendronephthya sp. Biochemistry (Mosc) 69, 901–908. Takeda, Y., Kawakami, H., Shigeta, S., Kamiya, H., Onizuka, R., Tanabe, K., Kato, Y., 2000. Purification of a novel allergen (SAP-1), which induces allergic asthma, from the red soft coral (Alcyonium gracillimum). Allergol. Int. 49, 213–218.
doi:10.1016/j.jbiotec.2008.07.1264 VI1-YP-026
References Engelmann, J., Volk, J., Leyhausen, G., Geurtsen, W., 2005. ROS formation and glutathione levels in human oral fibroblasts exposed to TEGDMA and camphorquinone. J. Biomed. Mater. Res. Part B: Appl. Biomater. 75B, 272–276. Jeon, Y.J., Kim, S.K., 2001. Effect of antimicrobial activity by chitosan oligosaccharide N-conjugated with asparagine. J. Microbiol. Biotechol. 11, 281–286. Mendis, E., Kim, M.M., Rajapakse, N., Kim, S.K., 2007. An in vitro cellular analysis of the radical scavenging efficacy of chitooligosaccharides. Life Sci. 80, 2118–2127.
doi:10.1016/j.jbiotec.2008.07.1265 VI1-YP-027 Antioxidative effect of methanolic extract of Sargassum thunbergii in macrophage RAW264.7 and HL60 Jung-Ae Kim 1 , Young-Sook Cho 1 , Won-Kyo Jung, Moon-Moo Kim, Se-Kwon Kim 1,∗ 1 Department of Chemistry, Pukyong National University, Busan 608737, Republic of Korea
Reactive oxygen species (ROS) are generated in all aerobic organisms. Imbalances in the production and removal of ROS have been hypothesized to play a causative role in numerous diseases such as cancer, neuronal degenerative diseases such as photoaging, atherosclerosis, and arthritis. Algae extracts containing a large amount of polyphenolic compounds have been studied as potential natural antioxidants in recent years. The aim of this study was to investigate potential antioxidant effects of marine algae on cultured macrophage RAW264.7 and HL60 cells. The brown algae Sargassun thunbergii is distributed widely coastal area of East Asia. In this study, the free radical scavenging effects of methanolic extract S. thunbergii (ST) on ROS production was investigated. The levels of intracellular ROS was measured using 2,7-dichlorofluorescindiacetate. Furthermore, membrane lipid peroxidation and protein oxidation inhibition was significantly decreased in a dose-dependent manner by treatment of ST extracts. Besides, the ST extract inhibited myeloperoxidase (MPO) activity which coverts H2 O2 into HOCl in human myeloid cell (HL60) and DNA oxidation caused by hydroxyl radical generated by Fenton reaction.
Antioxidant effect of novel chitooligosaccharides derivatives Dai-Nghiep Ngo 1,2,∗ , Moon-Moo Kim, Won-Kyo Jung, Sang-Hoon Lee, Tae-Kil Eom, Se-Kwon Kim 1 1
Marine Bioprocess Research Center, Pukyong National University, Busan 608-737, South Korea 2 Department of Biochemistry, Faculty of Biology, University of Sciences, VNU-HCMC, Ho Chi Minh City, Viet Nam E-mail address: [email protected] (D.-N. Ngo).
The object of this research is to improve antioxidant effect of chitooligosaccharides (COS) by grafting some phenolic acid such as cinnamic acid, coumaric acid and ferulic acid onto amino groups of COS (Jeon and Kim, 2001). The antioxidant effect of novel COS
References Li, C.W., Ja, A.A., In, W.I., 2005. Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts. J. Agric. Food Chem. 53, 4207–4212. Rajapakse, N., Kim, M.M., Kim, S.K., 2007. Inhibition of free radical-mediated oxidation of cellular biomolecules by carboxylated chitooligosaccharides. Bioorg. Med. Chem. 15, 997–1003.
doi:10.1016/j.jbiotec.2008.07.1266